Cloning, expression, crystallization and preliminary X-ray data analysis of norcoclaurine synthase from Thalictrum flavum

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 Apr 1;64(Pt 4):281-3. doi: 10.1107/S1744309108005678. Epub 2008 Mar 21.

Abstract

Norcoclaurine synthase (NCS) catalyzes the condensation of 3,4-dihydroxyphenylethylamine (dopamine) and 4-hydroxyphenylacetaldehyde (4-HPAA) as the first committed step in the biosynthesis of benzylisoquinoline alkaloids in plants. The protein was cloned, expressed and purified. Crystals were obtained at 294 K by the hanging-drop vapour-diffusion method using ammonium sulfate and sodium chloride as precipitant agents and diffract to better than 3.0 A resolution using a synchrotron-radiation source. The crystals belong to the trigonal space group P3(1)21, with unit-cell parameters a = b = 86.31, c = 118.36 A. A selenomethionine derivative was overexpressed, purified and crystallized in the same space group. A complete MAD data set was collected at 2.7 A resolution. The model is under construction.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbon-Nitrogen Ligases / biosynthesis
  • Carbon-Nitrogen Ligases / chemistry*
  • Carbon-Nitrogen Ligases / genetics*
  • Cloning, Molecular
  • Crystallization
  • Crystallography, X-Ray
  • Gene Expression Regulation, Plant / physiology*
  • Plant Proteins / biosynthesis
  • Plant Proteins / chemistry*
  • Plant Proteins / genetics*
  • Thalictrum / enzymology*
  • Thalictrum / genetics*

Substances

  • Plant Proteins
  • Carbon-Nitrogen Ligases
  • norcoclaurine synthase